Background: The current approaches to distinguish follicular adenomas (FA) and follicular thyroid cancer (FTC) at the pre-operative stage have low predictive value. Liquid biopsy-based analysis of circulating extracellular vesicles (EVs) presents a promising diagnostic method. However, the extreme heterogeneity of plasma EV population hampers the development of new diagnostic tests. We hypothesize that the isolation of EVs with thyroid-specific surface molecules followed by miRNA analysis, may have improved diagnostic potency. Methods: The total population of EVs was isolated from the plasma of patients with FA (n = 30) and FTC (n = 30). Thyroid peroxidase (TPO)-positive EVs were isolated from the total populations using immune-beads. The miRNA from the TPO(+)EVs obtained from the plasma of FA and FTC patients was assayed by RT-PCR. The diagnostic potency of the selected miRNAs was estimated by the receiver operating characteristic (ROC) analysis. Results: TPO(+)EVs can be efficiently isolated by immunobeads. The analysis of Let-7 family members in TPO(+)EVs allows one to distinguish FA and FTC with high accuracy (area under curve defined by ROC = 0.77–0.84). Conclusion: The isolation of TPO(+)EVs, followed by RT-qPCR analysis of Let-7 family members, may present a helpful approach to manage follicular nodules in the thyroid gland.

Abstract Background and aims : In Risk Classifier for Endometrial Cancer/EC (ProMisE) 4 molecular biological types of this tumor are described recently [1,2,3] and need an additional research, including evaluation of hormone-associated characteristics of both tumor tissue and patients [4]. Aim of the work was a study of estrogen (ER) and progesterone (PR) receptors in comparison with CD68+ macrophage infiltration (which promotes the invasion of tumor cells into the myometrium [5]) and expression of MDM2 protein, a negative regulator of p53 [6], in EC types presented in the ProMisE classification. Materials and methods: The tumor tissue of 218 EC patients included in the study (mean age 60.6 years) was assigned according to the data of genetic and immunohistochemical analysis to the types of carcinomas with gene POLE mutations, deficiency of mismatch repair proteins (MMR-D), expression (positive or diffuse) of p53 oncoprotein and to the type without characteristic molecular profile, WCMP. Immunohistochemistry was used also for evaluation of ER (Ventana antibodies, clone SP1) and PR (Ventana antibodies, clone 1E2) according to Allred, MDM2 (antibodies ABCAM, dilution 1:200) and macrophage marker CD68 (DAKO antibodies, clone CD8/144B). Results: According to the averaged data, the highest expression of ER and PR was found in EC types MMR-D and WCMP, and the lowest, respectively, in types POLE and p53+. Most often, positive expression of MDM2 (in 93.2% and 96.9% of studied cases) was detected respectively in MMR-D and p53+ type of EC, indicating, therefore, a positive relationship between MDM2 and the presence of steroid receptors in the first of these types (MMR-D) and negative - in the second of them (p53+). Expression of CD68+ macrophages demonstrated (contrary to the EC types POLE and p53+) a tendency to the lower values in types MMR-D and WCMP (128.0 ± 8.1 and 113.5 ± 6.3 cond.un.), i.e. in tumors with potential sensitivity to estrogen. Conclusions: The results indicate the importance of taking into account of both - the molecular biological type of the EC as well as the role of microenvironment of tumor cells, including the colonization of the neoplasm’ tissue by macrophages (and possibly lymphocytes) and the features of hormonal signal transmission in it. For further analysis, it is desirable to consider also the EC histotype, as one of the factors underlying various prognostic groups of this tumor [7]. References: 1.Talhouk A, McAlpine JN. Gynecol Oncol Res Pract. 2016; 3:14. 2. Talhouk et al., Cancer. 2017; 123(5):802–813. 3. Kommoss FK et al., Br J Cancer. 2018;119(4):480–486. 4. Berstein et al. Future Oncol. 2019; 15(12):1335–1346. 5. Jing X. et al. Immunol Cell Biol. 2019; 97(6):563–576. 6. Zou X. et al. Medicine (Baltimore).2018;97(49):e13273. 7. Bosse T. et al. Am J Surg Pathol. 2018 May;42(5):561–568.

Aim: To compare endocrine characteristics of endometrial cancer (EC) patients based on recent molecular EC types classification. Materials & methods: A total of 234 treatment-naive EC patients as well their tumors were studied. Results: Patients with POLE mutations demonstrated tendency to lower body mass index (BMI) and higher serum estradiol. Patients with p53 overexpression were older and had higher diabetes incidence. In the without characteristic molecular profile group there was no difference in fasting serum insulin, estradiol and testosterone levels between women with BMI >= 30.0 and <30.0. The mismatch repair deficient group patients had a tendency toward later menarche compared with the without characteristic molecular profile group one. Conclusion: Studied endocrine characteristics are associated with BMI or tumor molecular-biological type that might be relevant to EC genesis, course and prognosis.

Aim. To study serum content of a number of known adipokines, myokines and cytokines and compare obtained data with overweight phenotype and characteristics of tumor in untreated patients with endometrial cancer (EC) Materials and methods. The study included 88 patients with a mean age 60.080.67 years and a mean body mass index (BMI) 32.900.83. Patients were subjected to anthropometry and analysis of laboratory parameters, including serum level of leptin, adiponectin, omentin, prefilin (Pref-1), myostatin, irisin, IL-6 as well as insulinemia and insulin resistance index (HOMA-IR). On the basis of combination of anthropometric and laboratory data patients with overweight (BMI 25.0) were subdivided into the groups with "standard" (S) and conditionally "metabolically healthy" (MH) phenotype. Results. Levels of leptin, insulin and adiponectin in the serum of patients with EC are associated with BMI value and demonstrate significant differences between S and MH groups. Levels of prefilin, myostatin, and IL-6 are not associated with an increase in BMI, but are also different in patients with S and MH phenotype of overweight.For levels oа irisin, omentin, and TNF-alphathere is no peculiar dependence both, the BMI, and of belonging to a group with S or MH phenotype. Omentin level in the serum is associated with less favorable tumor differentiation (MH group), while IL-6 level with a more advanced stage of the tumor (all patients and group S). Conclusion. Adipokines, myokines and cytokines circulating in blood of EC patients vary in their connections with BMI or with "standard" or "metabolically healthy" phenotype of its excess. They vary also in relation to EC features, which in sum may have practical importance.

Purpose Adipose tissue products may contribute to endometrial cancer (EC) initiation and further growth that encourages the analysis of this issue in patients with different obesity phenotypes. Methods/patients Omental fat depot characteristics were studied in EC patients (n = 57) with “standard” (SO) or “metabolically healthy” (MHO) obesity. Collected omental samples were evaluated by immunohistochemistry /IHC/ for brown fat marker UCP1, CYP19 (aromatase) and macrophage infiltration markers (CD68, CD163, crown-like structures/CLS) expression. Total RNA extracted from the same samples was investigated for UCP1, CYP19, PTEN and adipokine omentin mRNA. Results Immunohistochemistry data revealed a statistically significant increase in aromatase and CD68 expression and tendency to increase of UCP1 expression in SO patients’ omental fat compared to samples obtained from MHO patients. Additionally, positive correlation of EC clinical stage with UCP1 protein and its mRNA content in omental fat was pronounced in MHO as well as SO group, while with omentin mRNA it was discovered only in patients with SO. An inclination to the correlation with better tumor differentiation was seen for UCP1 and CD68 protein expression in patients with MHO and with worse (high grade) differentiation—for CD68 expression in the group with SO. Conclusions In aggregate, this suggests that obesity phenotype has significant impact on omental fat tissue characteristics which is related to the clinical course of EC and may have practical consequences.

Thyroid cancer (TC) is the most common endocrine malignancy and its incidence has increased over the last few decades. As has been revealed by a number of studies, TC tissue’s micro-RNA (miRNA) profile may reflect histological features and the clinical behavior of tumor. However, alteration of the miRNA profile of plasma exosomes associated with TC development has to date not been explored. We isolated exosomes from plasma and assayed their characteristics using laser diffraction particle size analysis, atomic force microscopy, and western blotting. Next, we profiled cancer-associated miRNAs in plasma exosomes obtained from papillary TC patients, before and after surgical removal of the tumor. The diagnostic value of selected miRNAs was evaluated in a large cohort of patients displaying different statuses of thyroid nodule disease. MiRNA assessment was performed by RT-qPCR. In total, 60 patients with different types of thyroid nodal pathology were included in the study. Our results revealed that the development of papillary TC is associated with specific changes in exosomal miRNA profiles; this phenomenon can be used for differential diagnostics. MiRNA-31 was found to be over-represented in the plasma exosomes of patients with papillary TC vs. benign tumors, while miRNA-21 helped to distinguish between benign tumors and follicular TC. MiRNA-21 and MiRNA-181a-5p were found to be expressed reciprocally in the exosomes of patients with papillary and follicular TC, and their comparative assessment may help to distinguish between these types of TC with 100 % sensitivity and 77 % specificity.

AIM:The goal of this study was to determine if the single nucleotide polymorphisms marking potential sensitivity to metformin (MF) correlate with hormone-metabolic status as well as with actual response to MF in postmenopausal cancer patients with or without Type 2 diabetes mellitus and in diabetics without cancer.PATIENTS & METHODS:The carriage of ten different SNPs was evaluated in all patients by PCR, and hormone-metabolic status was estimated by anthropometry, ELISA and enzyme colorimetric assays. The response to daily 1-1.7 g of MF was studied based on hormone-metabolic parameters and indirect end points (endometrium thickness, mammographic breast density).RESULTS & CONCLUSION:The changes in evaluated 'antineoplastic' and metabolic response marker values were seen in 33.3 and 61.8% of the cases, respectively. Several genetic markers were found that showed an inclination to less frequent 'antineoplastic' or more frequent metabolic response to MF which may be helpful in further studies of this drug in cancer patients.

Background: As endometrial cancer (EC) prevalence increases with obesity, we aimed to determine whether EC characteristics depend upon obesity type: ‘standard’ (SO) or ‘metabolically healthy obesity’ (MHO). Patients & methods: 258 EC patients were included. Data on anthropometry, blood hormones, lipids and glucose, and tumor features were collected. Results: EC clinicopathologic characteristics and clinical stage correlate differently with BMI and obesity type. BMI is related inversely with tumor grade while SO patients are characterized by a more advanced clinical stage than those with MHO. Besides typical insulin resistance signs, EC patients with SO often display a higher serum leptin/adiponectin ratio compared with MHO patients. Historical data suggest a gradual increase in EC patient height and weight, and a decrease in MHO prevalence. Conclusion: It is currently unknown whether the latter observation reflects the evolution of EC, or obesity alongside the current epidemic. Regardless, the reduced MHO prevalence demonstrates the need for more intensive preventive measures aimed at obesity and obesity-associated conditions, including different EC subtypes.

Aim & methods: The present pilot study included 50 differentiated thyroid cancer (DTC) patients (mean age: 45.8 +/- 1.8 years, range: 18-73 years) who were followed after surgery for in average 30.0 +/- 2.6 months. All patients were subdivided into two groups as having either <2 ng/ml or >= 2 ng/ml blood thyroglobulin level 3 weeks after the operation (3-WTT). Results: Subsequent tumor progression was revealed more often in patients with higher thyroglobulinemia (>= 2 ng/ml) in both low-and high-risk DTC groups. Patients with high-risk DTC and 3-week thyroglobulin levels >= 2 ng/ml were more likely to have a higher pre-surgical thyrotropin (TSH) levels. On the contrary, patients with low-risk DTC and 3-week thyroglobulin level >= 2 ng/ml demonstrated tendency to higher preoperative serum insulin levels and higher BMI. Conclusion: These differential findings could suggest that, whereas maximal suppression of TSH is reasonable in high-risk DTC patients, in low-risk DTC patients, for whom this is not justified, a moderate TSH suppression supplemented by multivalent drugs, such as antidiabetic biguanide metformin, could be advised.

Metformin is a well-known antidiabetic medication, which, besides diabetes, may be involved into modulation of other age-related pathologies, including cancer. The study concerns 12 gene polymorphisms divided into 2 groups consisting of 6 genes each. The first group was composed from so-called "standard" (S) polymorphisms, for which the connection with metabolic response to metformin is already established. The second group included polymorphisms of genes encoding proteins possibly connected with diabetes mellitus type 2 (DM2), impaired glucose tolerance or cancer and entitled here as "associated" (A). A total of 156 postmenopausal women (average age 60.7 ± 0.7) were included, 37 of them healthy, 64 with type DM2 and concurrent treatment-naïve cancer (mostly breast, endometrial or colorectal cancer), 32 with DM2 without cancer, and 23 with treatment-naïve cancer and normal glucose tolerance. The leading metformin response S-marker in combined group of DM2 patients was the CC variant of OCT1-R61C polymorphism of organic cation transporter protein 1 gene. In cancer patients without DM2, this position belonged to AC and AA genotypes of OCT1_rs622342 polymorphism. Among the A-polymorphisms, GA variant of sex hormone-binding globulin gene SHBG_D356N was less frequently observed in DM2 patients with or without cancer. Besides, in diabetics, the same polymorphic variant of SHBG as well as GC genotype of oxidized lipoprotein receptor OLR1_G501C and GG genotype of locus rs11065987 near BRAP gene were carried rather often in combination with "metformin-positive" variant of OCT1_R61C. In addition, carriers of OCT1_R61C and OCT1_rs622342 polymorphisms with potentially positive reaction to metformin had higher insulin resistance score (HOMA-IR) values. Received data lead to the conclusion that postmenopausal diabetics, both with and without cancer, differ in genetic stigmata of potential response to metformin less than they differ from cancer patients without DM2. As genetic polymorphisms associated with metabolic and anticancer metformin (and, possibly, phenformin) effects may be different, this subject requires further investigation.

The goal of this study was to identify a set of fluorescence in situ hybridization probes for the detection of dysplasia and adenocarcinoma in patients with Barrett's esophagus. We examined 170 brushing specimens from 138 patients with Barrett's esophagus or a history of Barrett's esophagus using fluorescence in situ hybridization with probes to 5p15, 5q21-22, centromere 7, 7p12, 8q24.12-13, centromere 9, 9p21, centromere 17, 17p13.1, 17q11.2-12, 20q13.2, and centromere Y. Receiver-operator curves were used to determine the sensitivity and specificity of various four-probe combinations for detecting low-grade dysplasia, high-grade dysplasia, and esophageal adenocarcinoma. Endoscopic biopsy results were used as the gold standard. Numerous four-probe combinations provided a similarly high sensitivity and specificity. Of these, a set consisting of probes to 8q24, 9p21, 17q11.2, and 20q13.2 was found to have a sensitivity and specificity, respectively, of 70% and 89% for low-grade dysplasia, 84% and 93% for high-grade dysplasia, and 94% and 93% for esophageal adeno-carcinoma. This probe set was chosen for future prospective clinical evaluations based on its high sensitivity and specificity, its ability to distinguish adenocarcinoma and high-grade or low-grade dysplasia from lesser diagnostic categories, and the favorable signal quality for each of the probes.

Aromatase activity (AA) was evaluated totally in 80 tumors collected from primary endometrial cancer (EC) patients. All patients were divided into cases belonging to the types I or II of EC (respectively, 50 and 30 observations). Samples of malignant endometrium from type II demonstrated inclination to the higher AA in comparison with type I samples; the difference reached level of statistical significance in non-smoking patients (p=0.02). Although no positive correlation was revealed between AA in EC tissue and percentage of cells with DNA damage in normal endometrium from the same patients, the rate of DNA damage (percent of comets, comet's tail average length, etc.) was higher in intact endometrium collected from patients with type II of the disease. In 19 tumor samples, CYP19 gene expression was evaluated by RT-PCR and level of mRNA signal demonstrated positive correlation with AA (R(s)=+0.63, p=0.05) in the whole this material. Of note, though, CYP19 mRNA expression was not revealed in six cases, and all of them belonged to the type I of disease. Finally, in 23 EC patients (15 with type I and 8 with type II of the disease) effects of 2 weeks treatment with letrozole (10 pts) and exemestane (13 pts) were evaluated in neoadjuvant setting. Although diminishing of endometrial M-echo signal and the increases in FSH and LH concentration after treatment were more pronounced in type I patients, decrease in tumor PR content (p=0.04) was more revealing in patients with type II of EC; besides, the decreases in AA in tumor tissue by the end of treatment were noted predominantly in patients with lower body weight (BMI <27.5). Thus, although type II of EC is frequently considered as hormone-independent, increased ability of this type of the tumor to estrogen biosynthesis (at CYP19 gene and protein level) may lead to the reconsideration of such conclusion and warrants further investigation. The search of possible ethnic differences in AA and in the biologic response to aromatase inhibitors in EC can be of importance too.

Among the factors for estrogen and progesterone receptors (ER and PR) negativity of tumors of reproductive tissue special attention., is attracted by ability of the tumor produce estrogens (as intratumoral regulators of ER and PR) through reaction of aromatization. 101 samples of tumor tissue (64 cases of breast cancer and 37 cases of endometrial carcinomas) mostly from postmenopausal women were studied. When combined group of the tumors was divided on the basis of receptor-negativity or positivity (with cut-point correspondingly less than or equal to and >10 fM/mg protein) among samples with higher aromatase activity (>7 fM/mg protein/h), a tendency to higher number of ER-negative tumors was found (p=0.07). Such association was significant for breast (p=0.04) but not for endometrial cancer (p>0.5) samples. No evidence of PR contents dependence of tumor aromatase activity was revealed which may be connected with disturbance of estrogen signal transfer. Thus, inverse relation between steroid receptor levels and aromatase activity may be tissue- and receptor type-dependent.

Peculiarities of the estrogens influence on target tissues is one of the crucial problems in understanding of the estrogen-induced carcinogenesis and anticarcinogenesis mechanisms. Conditions or factors enhancing the genotoxic component in total effect of estrogens (on the uterine tissue, in particular) are very important, since these factors may influence both the hormonal carcinogenesis type and biological properties of the developing hormone-dependent tumors. In this study female rats (3 months of age at the beginning of experiment) have been given plain water (group 1) or 5% ethanol solution over 4 months. Rats which received ethanol were further divided into 6 groups (groups 2-7). During last 2 months of the experiment N-acetylcysteine was given to rats in group 3, ascorbic acid (vitamin C) and alpha-tocopherol (vitamin E)--to group 4, melatonin--to group 5, carnosine--to group 6; the rats in group 7 swam for 5 days a week according to the so called developing schedule. 2.5 weeks before the end of experiment all rats underwent bilateral ovariectomy, and over 11 days preceding the last day of the experiment they received injections of estradiol (2 microg intramuscularly daily). When the experiment was over, estradiol and cholesterol blood levels, progesterone receptors content, peroxidase activity, proliferation index, percent of cells in S and G2/M phases, thickness of endometrium and rate of DNA damage in uterine tissue (COMET assay) and estradiol 2-hydroxylase activity in liver tissue were measured. The conclusion was that administration of 5% ethanol combined with estrogen injections results in genotoxic (G) changes in the uterus, which may be prevented by giving N-acetylcysteine or melatonin. Combination of vitamins C and E enhances some features of hormonal (H) estrogen effects (uterine weight, induction of progesterone receptors), but attenuates the other (proliferation index). Consequently, the combination of N-acetylcysteine and optimal doses of ascorbic acid and alpha-tocopherol may be recommended for prevention of the phenomenon of switching of estrogen effects [PSEE] (e.g. enhancement of G-component and decrease of H-component), observed particularly in cases of the treatment with tobacco smoke or ethanol consumption in more than moderate (15%) concentrations, which lead to the increased risk of genotoxic type of hormonal carcinogenesis.

OBJECTIVE:To investigate the short-term hormonal and clinical effects of the aromatase inhibitor letrozole (Femara) in patients with endometrial cancer. MATERIALS AND METHODS:Ten previously untreated, post-menopausal patients (mean age 59 years) with endometrial cancer, predominantly stage I disease, received letrozole 2.5mg per day for 14 days before surgery. Clinical, sonographic, morphologic, cytologic, and hormonal-metabolic parameters (blood estradiol, follicle-stimulating hormone (FSH), luteinizing hormone (LH), glucose, and cholesterol by radioimmunoassay, enzyme immune assay, or enzyme-colorimetric methods; tumor progesterone receptors by ligand-binding assay; and aromatase activity by 3H-water release assay) were evaluated before and after treatment. RESULTS:Treatment was well-tolerated in all patients. In two patients, pain relief in the lower part of the belly and/or decrease in intensity of uterine discharge was reported. In the three cases, substantial decreases in endometrial M-echo (ultrasound) signal were noted; the mean value of this parameter after treatment was 31.1% lower than before treatment. Blood estradiol concentration decreased by an average of 37.8% after letrozole therapy, and tumor progesterone receptor levels and aromatase activity decreased by 34.4 and 17.5%, respectively. Treatment with letrozole did not influence surgery. CONCLUSIONS:These data show that short-term treatment with letrozole in the neoadjuvant setting resulted in some positive clinical changes. Longer-term and larger-scale trials of neoadjuvant letrozole in endometrial cancer are warranted.